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A shotgun approach to identify mechanical nociception genes

Melissa G. Christianson, Stephanie E. Mauthner, W. Daniel Tracey
doi: https://doi.org/10.1101/077644
Melissa G. Christianson
1Department of Neurobiology, Duke University Medical Center, Durham, NC 27710
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Stephanie E. Mauthner
1Department of Neurobiology, Duke University Medical Center, Durham, NC 27710
2Gill Center for Biomolecular Sciences and Department of Biology, Indiana University, Bloomington, Indiana, 47405
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W. Daniel Tracey
1Department of Neurobiology, Duke University Medical Center, Durham, NC 27710
2Gill Center for Biomolecular Sciences and Department of Biology, Indiana University, Bloomington, Indiana, 47405
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  • For correspondence: dtracey@indiana.edu
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Abstract

The molecular mechanisms of sensing noxious mechanical force by nociceptive sensory neurons remain poorly understood. Traditional methods for probing mechanical nociception behavioral responses are labor intensive and involve the testing of one animal at a time. This time consuming process of behavioral testing has largely precluded large scale analyses. Indeed, large scale genetic screens that have been performed thus far have been largely restricted to the investigation of ion channel genes [1]. Here we describe a new behavioral assay for mechanical nociception in which tens of animals can be stimulated simultaneously. In this assay, third instar larvae of the genetically tractable organism Drosophila melanogaster are mechanically stimulated with tungsten particles that are fired from a gun. We have used the new assay to carry out a genetic screen in which we investigated the function of 231 nociceptor enriched genes with tissue-specific RNA interference. Targeting of 21 genes resulted in mechanically insensitive phenotypes and targeting of a single gene resulted in a hypersensitive mechanical nociception phenotype. Six of the identified genes were previously uncharacterized and these were named after famed Roman gladiators (Spartacus (CG14186), Commodus (CG1311), Flamma (CG10914), Crixus(CG6685), Spiculus (CG10932), and Verus (CG31324)).

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted October 18, 2016.
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A shotgun approach to identify mechanical nociception genes
Melissa G. Christianson, Stephanie E. Mauthner, W. Daniel Tracey
bioRxiv 077644; doi: https://doi.org/10.1101/077644
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A shotgun approach to identify mechanical nociception genes
Melissa G. Christianson, Stephanie E. Mauthner, W. Daniel Tracey
bioRxiv 077644; doi: https://doi.org/10.1101/077644

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